神经肽PACAP27抑制线粒体依赖性细胞凋亡途径对环磷酰胺所致大鼠睾丸损伤的改善作用  

Improvement effects of neuropeptide PACAP27 on cyclophosphamide-induced testicular injury in rats by inhibiting mitochondria-dependent apoptosis pathway

作　　者：廖源 王开举 李浩僡 陈惠萍 李选逸 黄勇[1] LIAO Yuan;WANG Kaiju;LI Haoyan;CHEN Huiping;LI Xuanyi;HUANG Yong(Department of Reproductive Medicine,Second Affiliated Hospital,Hainan Medical College,Haikou 570311,China)

机构地区：[1]海南医学院第二附属医院生殖科,海南海口570311

出　　处：《吉林大学学报（医学版）》2022年第5期1266-1275,共10页Journal of Jilin University：Medicine Edition

基　　金：海南省卫健委卫生计生行业科研项目(20A200022)。

摘　　要：目的:探讨垂体腺苷酸环化酶激活多肽(PACAP) 27对环磷酰胺所致大鼠睾丸损伤的作用,阐明其可能机制。方法:60只SD大鼠随机分为对照组、模型组、PACAP27组和左卡尼汀组,每组15只;除对照组外,其余各组大鼠采用环磷酰胺造模并给药。给药结束后第3天,称量大鼠体质量和睾丸质量,计算睾丸指数;采用放射免疫法检测各组大鼠血清中促卵泡激素(FSH)、黄体生成素(LH)和睾酮(T)水平,HE染色观察各组大鼠睾丸组织病理形态表现,酶联免疫吸附测定(ELISA)法检测各组大鼠睾丸组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)水平,TUNEL染色检测各组大鼠睾丸组织细胞凋亡率,Western blotting法检测各组大鼠睾丸组织中线粒体凋亡途径相关蛋白表达水平;提取睾丸组织线粒体,采用线粒体分离试剂检测各组大鼠睾丸组织中线粒体膜通透性转换孔(mPTP)开放程度,JC-1荧光染色检测各组大鼠睾丸组织线粒体膜电位。结果:与对照组比较,模型组大鼠体质量、睾丸质量和睾丸指数均降低(P<0.05),血清中FSH、LH和T水平降低(P<0.05),睾丸组织中SOD和CAT活性降低(P<0.05),MDA水平升高(P<0.05),睾丸组织发生明显病理损伤,细胞凋亡率升高(P<0.05),睾丸组织中细胞色素C(Cyt C)、B细胞淋巴瘤2相关X蛋白(Bax)、含半胱氨酸的天冬氨酸蛋白水解酶3 (caspase-3)和含半胱氨酸的天冬氨酸蛋白水解酶9 (caspase-9)蛋白表达水平升高(P<0.05),B细胞淋巴瘤2(Bcl-2)蛋白表达水平降低(P<0.05),mPTP开放程度增强(P<0.05),线粒体膜电位降低(P<0.05)。与模型组比较,PACAP27组和左卡尼汀组大鼠睾丸质量和睾丸指数均升高(P<0.05),血清中FSH、LH和T水平升高(P<0.05),睾丸组织中SOD和CAT活性升高(P<0.05),MDA水平降低(P<0.05),睾丸组织病理损伤得到改善,细胞凋亡率降低(P<0.05),睾丸组织中Cyt C、Bax、caspase-3和caspase-9蛋白表达水平降低(P<0.05),Bcl-2蛋白�Objective: To explore the effect of pituitary adenylate cyclase activating polypeptide(PACAP)27 on the cyclophosphamide-induced testicular injury in the rats,and to clarify its possible mechanism. Methods:Sixty SD rats were randomly divided into control group,model group,PACAP27group and L-carnitine group,with 15 rats in each group. Except control group,the rats in other groups were treated with cyclophosphamide to establish the models and administrited with drugs. On the 3rd day after the administration,the weights of rats and the testis were weighed,and the testis index was calculated;the levels of serum follicle-stimulating hormone(FSH),luteinizing hormone(LH)and testosterone(T)of the rats in various groups were measured by radioimmunoassay, HE staining was used to observe the pathomorphology of testis tissue of the rats in various groups;the activities of superoxide dismutase(SOD),catalase(CAT)and the levels of malondialdehyde(MDA)in testis tissue of the rats in various groups were detected by enzyme linked immunosorbent assay(ELISA)method,TUNEL staining was used to detect the apoptotic rates of testicular tissue of the rats in various groups,and Western blotting method was used to detect the expression levels of mitochondrial apoptosis pathway-related proteins. The testicular tissue mitochondria were extracted,the opening degrees of the mitochondrial permeability transition pore(mPTP) of the rats in various groups were detected with mitochondrionl isolation reagent,and JC-1fluorescent staining was used to detect the mitochondrial membrane potential. Results:Compared with control group,the body weight,testis weight and testis index of the rats in model group were decreased(P<0. 05),the levels of FSH,LH and T in serum were decreased(P<0. 05),the activities of SOD and CAT in testis tissue were decreased(P<0. 05),the level of MDA was increased(P<0. 05),the testis tissue had obvious pathological damage,and the apoptotic rate was increased(P<0. 05),the expression levels of cytochrome C(Cyt C),B-cell lymphoma-2 ass

关 键 词：睾丸损伤 环磷酰胺 垂体腺苷酸环化酶激活多肽27 线粒体依赖性凋亡途径 

分 类 号：R697.22[医药卫生—泌尿科学]